Broken into pieces: The challenges of determining sulfonated azo dyes in biological reactor effluents using LC-ESI-MS/MS analysis.

Most studies on the biodegradation of textile azo dyes use color as parameter for measuring the efficiency of degradation. Although widely employed, spectrophotometric methods are susceptible to the interference of metabolites or degradation products from the biological treatment. We propose a method for determination of a model sulfonated azo dye (Direct Black 22, DB22) in wastewater using solid-phase extraction (SPE) and liquid chromatography - electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). MS analysis in negative electrospray ionization mode showed DB22 as the most abundant precursor ion, corresponding to [M-3Na + H]2-, which yields two radical anions of m/z 370.1 and m/z 645 after MS/MS fragmentation by collision-induced dissociation (CID). Calibration curve presented adequate linearity and precision in the range of 120-1500 ng mL-1, and recovery and detection limit were appropriate to the typically employed working concentrations. Nevertheless, we observed that standard heating of DB22 under alkaline conditions to simulate the production of wastewater during dye-baths resulted in loss of MS/MS signal, without affecting color. Further analysis showed that DB22 undergoes hydrolysis and does not remain unaltered in solution. Alternative methods of hydrolysis evaluated resulted in no MS/MS signal as well. SPE-LC-ESI-MS/MS analysis evidenced the structural change of DB22 in aqueous solution while the dyeing-capacity was preserved. This technique has also the potential of being tailored to consider the detection of the hydrolyzed fragments of azo dyes in wastewater for appropriate quantification, but it was not the scope of the current step of this research. Color remains as a more reliable parameter for monitoring azo compounds which are unstable in aqueous solution, while a more robust and holistic method needs to be developed for the speciation of the DB22 products of thermal hydrolysis.

Effect of Desensitizing Agents on the Bond Strength of Mild and Strong Self-etching Adhesives.

BACKGROUND: Desensitizing agents are used, almost as routine practice, in many adhesive restorative procedures. There is still debate as to their effect in dentin bonding, particularly with self-etching adhesives. The present study aimed to evaluate the effect of different desensitizing agents on the bond strength of mild and strong self-etching adhesive systems to dentin. MATERIALS AND METHODS: One hundred twenty recently extracted, noncarious human molars were used to obtain superficial dentin substrate for bonding. No desensitizer was used in the control groups. The experimental groups were pretreated with Gluma Desensitizer, MicroPrime B, and Dentin Desensitizer immediately prior to bonding with self-etching adhesives Optibond XTR, Xeno IV, and iBond. A bonding jig was used to fabricate composite cylinders, which were stored for either 24 hours or three months, after which the shear bond strength (SBS) was evaluated using a notched-edge testing device at a crosshead speed of 1 mm/min. Failure mode distribution was also evaluated at 24 hours and three months. A two-way analysis of variance, Tukey test, and Student t-test, with a significance level of p<0.05, were used for data analysis. RESULTS: At 24 hours, there was no significant difference in SBS when the same adhesive was used with any of the experimental desensitizing agents compared with the control group without desensitizer. However, at three months, Dentin Desensitizer bonded with Optibond XTR demonstrated significantly lower SBS (p<0.001), while Gluma bonded with iBond showed significantly higher SBS values (p=0.034) relative to their corresponding control group. Only MicroPrime B bonded with Xeno IV and iBond with no desensitizer demonstrated a significant reduction in SBS after three months (p=0.034 and p=0.002, respectively). The most prevalent type of failure was adhesive. CONCLUSION: Desensitizing agents can be used in combination with self-etching adhesives to control hypersensitivity without adversely affecting their bond strength to dentin.

Transdentinal cytotoxicity of carbodiimide (EDC) and glutaraldehyde on odontoblast-like cells.

OBJECTIVE: To evaluate the transdentinal cytotoxicity of three different concentrations of carbodiimide (EDC) or 5% glutaraldehyde (GA) on MDPC-23 cells. METHODS: Seventy 0.4-mm-thick dentin disks obtained from human molars were adapted to artificial pulp chambers. MDPC-23 cells were seeded on the pulpal surface of the disks. After 48 hours, the occlusal dentin was acid-etched and treated for 60 seconds with one of the following solutions (n=10): no treatment (negative control); 0.1 M, 0.3 M, or 0.5 M EDC; 5% GA; Sorensen buffer; or 29% hydrogen peroxide (positive control). Cell viability and morphology were assessed by methyltetrazolium assay and scanning electron microscopy (SEM), respectively. The eluates were collected after the treatments and applied on MDPC-23 seeded in a 24-well plate to analyze cell death, total protein (TP), and collagen production. The last two tests were performed 24 hours and seven days after the challenge. Data were analyzed by Kruskal-Wallis and Mann-Whitney tests (p<0.05). RESULTS: EDC at all test concentrations did not reduce cell viability, while 5% GA did increase cell metabolism. Cell death by necrosis was not elicited by EDC or 5% GA. At the 24-hour period, 0.3 M and 0.5 M EDC reduced TP production by 18% and 36.8%, respectively. At seven days, increased TP production was observed in all groups. Collagen production at the 24-hour period was reduced when 0.5 M EDC was used. After seven days, no difference was observed among the groups. SEM showed no alteration in cell morphology or number, except in the hydrogen peroxide group. CONCLUSIONS: Treatment of acid-etched dentin with EDC or GA did not cause transdentinal cytotoxic effects on odontoblast-like cells.

Color Stability Behavior of Methacrylate-based Resin Composites Polymerized with Light-emitting Diodes and Quartz-Tungsten-Halogen.

UNLABELLED: Despite significant developments in improving the optical properties of resin composite materials, their color stability remains a challenge. This study aimed to evaluate the shade stability of light-polymerized, methacrylate-based resin composites with different filler particle composition (microfill, minifill, nanohybrids, and microhybrids) polymerized with quartz-tungsten-halogen (QTH) and light-emitting diodes (LED). METHODS AND MATERIALS: Composite discs were fabricated from Tetric EvoCeram, Premise, Artiste, and Beautifil II (nanohybrids); Filtek Supreme Plus and Vit-l-escence (microhybrids); Heliomolar (microfill); and Estelite Sigma Quick (minifill) using a Teflon mold. The specimens were irradiated either with QTH (Elipar 2500; 600 mW/cm(2)) for 40 seconds or with LED (Bluephase G2; 1200 mW/cm(2)) for 20 seconds. Color parameters were measured with a colorimeter before and after polymerization and at 24 hours, one week, one month, and three months. Color change was calculated among the different storage periods. RESULTS: There was a significant effect of the composite, time, and their interaction (p<0.001) but no effect of the polymerization unit on the color stability. Color changes immediately after polymerization and at 24 hours (4.22 and 3.88 for LED; and 4.08 and 3.82 for QTH) were not significantly different from each other but were both significantly higher than changes after one week (0.96 and 0.78), one month (1.12 and 1.02), and three months (1.27 and 1.11) for LED and QTH, respectively (p<0.001). CONCLUSIONS: Color changes were observed for all the materials that were dependent on the type of composite but not on the polymerization unit. These color shifts took place primarily immediately after polymerization and after 24 hours and were additive in nature.

Effect of post-thaw addition of seminal plasma on motility, viability and chromatin integrity of cryopreserved donkey jack (Equus asinus) spermatozoa.

Pregnancy rates in donkeys after artificial insemination with cryopreserved semen are still low, compared to the horse species. Addition of autologous seminal plasma to frozen-thawed semen appeared to improve pregnancy rates. The aims of this study were to evaluate (1) sperm motility and plasma membrane integrity after thawing (T0) and after one and 2 h (T1 and T2) of post-thaw incubation in either 0% (SP0) or 70% (SP70) autologous seminal plasma and (2) sperm motility, plasma membrane integrity and DNA quality (%COMP-αt) after thawing (T0) and after 2 and 4 h (T2 and T4) of post-thaw incubation in either 0% (SP0), 5% (SP5) or 20% (SP20) homologous seminal plasma. In experiment 1, seminal plasma decreased total and progressive sperm motility and plasma membrane intact spermatozoa immediately after dilution and at all following time points (p < 0.05). In experiment 2, total and progressive motility did not differ between treatments immediately after dilution and between SP0 and SP5 at T2, while they were lower in both SP5 and SP20 than in SP0 at T4. Plasma membrane intact sperm cells did not differ between SP0 and SP5 and were lower in SP20 at all time points. DNA quality was not affected by treatment immediately after dilution and was significantly worse for SP20 after 4 h of incubation (p < 0.05). The post-thaw addition of seminal plasma at the tested concentrations did not improve donkey frozen semen characteristics in vitro over time.

In vitro evaluation of benzalkonium chloride in the preservation of adhesive interfaces.

UNLABELLED: Inhibition of endogenous dentin matrix metalloproteinases (MMPs) by benzalkonium chloride (BAC) decreases collagen solubilization and may help improve resin-dentin bond stability. OBJECTIVE: This study evaluated the resin-dentin bond stability of experimental adhesive blends containing BAC and the stability of dentin matrices by assessing the mass loss and collagen solubilization from dentin beams pretreated with BAC. MATERIALS AND METHODS: Twenty-five healthy molars were used for the bond strength evaluation of a two-step etch-and-rinse adhesive (Adper Single Bond Plus, SB) modified with BAC or not. The following groups were tested: 1) SB with no inhibitor (control); 2) topical 2.0% chlorhexidine + SB; 3) 1.0% BAC etchant + SB; 4) 0.5% BAC-SB; and 5) 1.0% BAC-SB. Microtensile bond strength (µTBS) and failure mode distribution under standard error of the mean were evaluated after 24 hours and six months of storage in artificial saliva (AS). A two-way analysis of variance and Tukey test with a significance level of p<0.05 was used for data analysis. In addition, 30 completely demineralized dentin beams from human molars were either dipped in deionized water (DW, control) or dipped in 0.5% and 1.0% BAC for 60 seconds, and then incubated in AS. Collagen solubilization was assessed by evaluating the dry mass loss and quantifying the amount of hydroxyproline (HYP) released from hydrolyzed specimens after four weeks of incubation. RESULTS: The control group demonstrated lower µTBS than some of the experimental groups containing BAC at 24 hours and six months (p<0.05). When BAC was incorporated into the adhesive blend in concentrations of 0.5% and 1.0%, no reduction in dentin bond strength was observed after six months (p<0.05). Less mass loss and HYP release was seen for dentin matrices pretreated with BAC relative to the control pretreated with DW (p<0.05). CONCLUSION: This in vitro study demonstrates that BAC contributes to the preservation of resin-dentin bonds by reducing collagen degradation.

Effect of a chlorhexidine-containing adhesive on dentin bond strength stability.

PURPOSE: The present study aimed to investigate a novel adhesive system containing 0.2% chlorhexidine digluconate (CHX) for its ability to improve the stability of the adhesive interface compared with the use of 2% CHX as a therapeutic primer. Furthermore, the study aimed to confirm the inhibitory properties of these CHX concentrations (0.2% and 2.0%) on dentin matrix metalloproteinase activity by gelatin zymography. METHODS: Superficial dentin substrate for bonding was obtained from 120 non-carious human molars. A conventional adhesive Peak LC Bond and a CHX-containing adhesive Peak Universal Bond were used either in combination with 35% phosphoric acid (etch-and-rinse approach) or with self-etching primer (self-etch approach) for evaluation of the variables CHX treatment (2.0% therapeutic primer and 0.2% adhesive), adhesive approach (etch-and-rinse and self-etch), and storage time (24 hours and six months). A bonding jig was used to fabricate composite cylinders, which were stored for either 24 hours or six months, after which shear bond strength (SBS) was evaluated using a notched-edge testing device. A three-way analysis of variance and a Student t-test with a significance level of p<0.05 were used to analyze the data. Extracts from concentrated demineralized human dentin powder were subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis and incubated in the presence of 0.2% and 2.0% CHX. RESULTS: No significant effect of CHX treatment, adhesive approach, storage time variables, or their interactions on mean SBS was demonstrated (p<0.05). No significant difference between the control and the CHX-treated groups was detected for either adhesive technique at 24 hours or six months (p<0.05). No significant variation in mean SBS was detected after six months of storage (p<0.05). Zymographic analysis revealed bands of enzymatic activity for the group demineralized with phosphoric acid and complete inhibition of gelatinolytic activity for the groups treated with 0.2% and 2.0% CHX. CONCLUSIONS: CHX demonstrated inhibition of dentin proteolytic activity. However, when CHX was incorporated into a commercially available adhesive or used as a therapeutic primer, no difference in bond strength was observed at baseline or after six months of storage relative to the control group without CHX.

Clinical presentation of meningococcal disease in childhood.

Although relatively rare, meningococcal disease represents a global health problem being still the leading infectious cause of death in childhood with an overall mortality around 8%. Meningococcal meningitis is the most commonly recognized presentation, accounting for 80% to 85% of all reported cases of meningococcal disease (in half of these cases sepsis is also present concomitantly). The remaining 15-20% of cases are most commonly bloodstream infections only. Meningococcal serogroups A, B, and C account for most cases of meningococcal disease throughout the world. Recently, serogroups W-135 and X (predominantly in Africa) and group Y (in the United States and European countries) have emerged as important disease-causing isolates. Despite recent advances in medical management, the mortality rate of fulminant meningococcemia ranges from 15% to 30%. However, among survivors, 10-30% could have long term sequelae (i.e. sensoneural hearing loss, seizure, motor problems, hydrocephalus, mental retardation, and cognitive and behavioral problems). Considering the clinical severity of meningococcal disease, prevention represents the first approach for avoiding serious complications and possible deaths. The availability of new vaccines able to cover the emerging serotypes including A and Y as well as the availability on the market of new products that could prevent meningococcal B infection represent a great opportunity for the decrease of the burden of this complicated disease.

Donkey jack (Equus asinus) semen cryopreservation: studies of seminal parameters, post breeding inflammatory response, and fertility in donkey jennies.

The aims of this study were (1) to evaluate motility parameters of donkey jack (jack; Equus asinus) semen cryopreserved in INRA-96 (INRA; IMV Technologies, France, 2% egg-yolk enriched) using either glycerol (GLY) or ethylene glycol (EG) as a cryoprotector; (2) to compare in vitro the postthaw re-extension with homologous seminal plasma (SPL) or INRA; (3) to compare fertility in donkey jennies (jennies; Equus asinus) timed artificially inseminated with jack semen cryopreserved using GLY or EG, re-extended with INRA; (4) to compare fertility in jennies timed artificially inseminated with jack semen cryopreserved using GLY re-extended with SPL, INRA, or not re-extended (NN); and (5) to describe some preliminary results of the inflammatory uterine response postbreeding. Semen from two jacks was collected and frozen in an INRA-2% egg yolk extender added of either 2.2% GLY or 1.4% EG. Postthaw motility was evaluated by a computer-assisted motility analyzer. Uterine inflammatory response and fertility were evaluated after artificial insemination (AI) of 13 jennies with frozen-thawed semen, either further extended with INRA (Group GLY-INRA, 13 cycles, and EG-INRA, 8 cycles), or with SPL (Group GLY-SPL, 13 cycles), or not re-extended (GLY-NN, 5 cycles). In each cycle, jennies were bred twice with 500 × 10(6) sperm cells (250 × 10(6) from each jack), at fixed times after induction of ovulation, and uterus was flushed at 6 and 10 h after first and second breeding, respectively. Cells in the recovered fluid were counted and distinguished as polymorphonuclear neutrophils (PMN) or other cell types. Total and progressive motility did not differ between cryoprotectants, but were higher when semen samples were re-extended in INRA, compared with SPL (P < 0.05). Pregnancy was diagnosed by transrectal palpation and ultrasonography examinations at 14 and 16 days postovulation. In 7/13 (53.8%) jennies and 12/39 (30.4%) cycles postbreeding intrauterine fluid accumulation was observed, with no differences between treatments (P < 0.05). Polymorphonuclear neutrophil numbers and concentrations were higher in the first flushing compared with the second, and PMN concentration was higher in GLY-SPL than in GLY-INRA (P < 0.05). Pregnancy rates in GLY-SPL, GLY-INRA, EG-INRA, and GLY-NN were 8/13, 3/13, 2/8, and 1/5, respectively. There was no significant difference either between the two cryoprotectants re-extended in INRA, or between re-extension groups. There was however a trend for GLY-SPL to improve pregnancy rates compared with GLY-INRA (P = 0.055). These results indicate that it is possible to obtain similar postthaw sperm motility and pregnancy rates using GLY or EG as a cryoprotectant for donkey semen, and that in the conditions of this study the re-extension in SPL of thawed semen before AI showed a trend toward the improvement of fertility and increased PMN concentration in uterine flushings.

Can infants be protected by means of maternal vaccination?

The administration of vaccines is not usually recommended in pregnant women because of a fear of severe adverse events for the fetus. However, contraindication to vaccination applies only to vaccines based on live attenuated viruses for the theoretical possibility that they might infect the fetus. In contrast, the use of several inactivated vaccines is useful and recommended. As a result of the transplacental passage of antibodies, maternal immunization can reduce the risk of vaccine-preventable diseases that may occur in the first months of life before the start or completion of the suggested vaccination schedule. One of the best examples is vaccination against influenza that can protect pregnant women from a disease that can lead to hospitalization and death in a significantly higher number of cases than in the general population and can induce protective specific antibody levels as well as being effective in infants in the first months of life. Other examples are vaccinations against tetanus, pertussis, pneumococcal infections and Haemophilus influenzae type b infection. This review analyses the advantages and limitations of maternal immunization as revealed by experience and the main publications.

Cross-compatibility of methacrylate-based resin composites and etch-and-rinse one-bottle adhesives.

OBJECTIVE: To compare dentin shear bond strength (SBS) of four combinations of light-activated one-bottle adhesives and composites to determine if cross-compatibility exists, and to determine if the use of the same manufacturer's adhesive and composite results in higher SBS than systems that combine different manufacturers' products. METHODS: One hundred sixty human third molars were used for bonding (n=10). Specimens were treated with 37% phosphoric acid and one of four etch-and-rinse adhesives. Specimens were placed in a bonding jig, which was filled with one of four composites. Adhesives PQ1 (Ultradent), Excite (Ivoclar-Vivadent), Optibond Solo Plus (Kerr), and Single Bond (3M-ESPE) and composites Vit-l-Escence (Ultradent), Four Seasons (Ivoclar-Vivadent), Premise (Kerr), and Filtek Supreme Plus (3M-ESPE) were tested. SBS was measured at 24 hours and three months with a testing machine at a speed of 1 mm/min and expressed in MPa. A three-way analysis of variance and Tukey tests were used for data analysis. RESULTS: Significant differences were evidenced among composites for each adhesive system (p<0.001) and among adhesives for each composite system (p<0.001). Optibond Solo Plus and PQ1 yielded significantly higher bond strengths than Single Bond and Excite for all composite systems (p<0.05). All combinations, with the exception of two, demonstrated a decrease in bond strength values after aging. CONCLUSIONS: Cross-compatibility was demonstrated, indicating that etch-and-rinse one-bottle adhesive systems can be safely used with composites from different manufacturers without a compromise to the bond strength. Moreover, even higher mean SBS values were demonstrated for selective combinations of different manufacturers' products

[Quality control of defrosted cord blood units: results from an inter-laboratory study]. / Contrôle de qualité des greffons de sang placentaire décongelés : résultats d'une étude multicentrique.

PURPOSE: Today, haematopoietic stem cell graft from placental blood concerns more than 15 % of allogeneic grafts. An inter-laboratory study of the quality control of defrosted cord blood units has been coordinated by the French society for cell and tissue bioengineering (SFBCT), with the cord blood bank of Bourgogne Franche-Comté and controlled by the French health products safety agency (Afssaps). The aim of this study is to ensure the inter-laboratory reproducibility of the quality controls practised by the banks during defrosting. The cellular outputs were analyzed according to the defrosting techniques, according to the method used in flow cytometry: single-platform (SP) versus double-platform (DP), or the product nature, i.e. in total blood or miniaturized. METHODS: Forty-two units of placental blood (USP), which were out of range were provided for defrosting to 14 participating sites. USP were defrosted and controlled according to the procedures of each bank. Once the USP is defrosted, a part of the product was controlled by the site and the other part by Afssaps. Following controls were carried out: numeration of the total nucleated cells (TNC) and of CD34+ cells (made by a SP method in Afssaps) and functional assay. RESULTS: Concerning TNC, the defrosting sites obtained a cellular output of 94 %+/-28 in day 0 compared with an output of 72 %+/-24 in Afssaps showing a rather good stability of the USP transmitted with an average deviation of 23 %+/-22. The freezing process with or without reduction of volume does not affect this variation. Concerning the numeration of CD34+ cells, the average deviation between the participating sites and Afssaps was 29 %+/-23 compared with 21 %+/-16 for the sites using a SP method against 47 %+/-25 for those using a DP method. The CD34+ outputs are equal to 82 % +/- 60 in day 0 for the participating sites against 52 %+/-20 for Afssaps. For the sites using a DP method, it is stressed that this output is particularly high with a rate of 126 %+/-90 (n=15) whereas it is 62 %+/-20 (n=32) for the sites using a SP method. CONCLUSION: These results underline a good stability of viable CD34+ cells and a greater reliability of the SP methods for the CD34+ cell numeration for these defrosted USP. Lastly, the results of the functional assay regarding the average clonogenicities (equal to 15 %) reinforce the conclusions on the quality of the defrosted products.

Study of bone metabolism in patients with chronic HIV infection.

AIMS: Various studies have confirmed the high incidence of skeletal homeostasis modifications in subjects who are carriers of chronic HIV infections, and specific pharmacological treatments, which modify the metabolism and condition both the weight loss and the reshaping of the bones. The presence of a reduction in body mass index seems to contribute to the progressive deterioration of the skeletal framework. The aim of this study was to see whether the presence of HIV-seropositivity could constitute a risk factor for the development of osteoporosis/osteopenia, even in the light of the fact that our group was composed of patients with a concentrated age span well under the limit for both post-menopausal and senile osteoporosis, and with a median age superimposable for both sexes. MATERIALS AND METHODS: Our study involved 26 HIV+ patients with an average duration of infection equal to 6.7 +/- 4.8 years, and a range of seropositive duration between 6 months to 16 years. The prominent ultrasonometrical parameters are as follows: Broadband Ultrasound Attenuation, Speed of Sound, Stiffness Index or Quantitative Ultra-sound Index, Bone Mineral Density, and T-score. The biochemical study was carried out by assessing a marker of neoformation such as seric osteocalcine, and uninary pyridinoline and deoxipyridonoline as resorption markers. RESULTS: The results confirmed the presence of osteoporosis/osteopenia in 46% of the samples (11%, and 35%, respectively), with a progressive reduction in bone mineral density in relation to the duration of HIV infection. Assessment of the marker for bone metabolism showed a significant increase in osteocalcine in the female population compared to the males, without any significant variations in the normal values. CONCLUSIONS: Extreme variability in the morphological appearance at bone level during the course of HIV infection would lead us to believe that in the genesis of various forms, depending on the mechanisms and the time involved only in the parts defined, other attributable factors are responsible, not only for the progression of the core pathology and the possible interference of hormonal factors (behavioural and/or nutritional) directly correlated with the state of infection, but also for the dismetabolic effects of the antiretroviral drugs.

Evaluation of oxidative stress in rheumatoid and psoriatic arthritis and psoriasis.

INTRODUCTION: The aim of the present study is to discuss the importance of the processes of oxidative stress in the pathogenesis of certain autoimmune diseases, to search for an appropriate assessment marker, and to debate current approaches which have been proposed for the treatment of Rheumatoid Arthritis (RA), Psoriatic Arthritis (PsA), and Psoriasis (Ps). MATERIALS AND METHODS: The total antioxidant capacity (TAC), the thiolic capacity (TC), and the serum hydroperoxide concentration (SHC) were measured in 37 subjects: 13 with RA, 8 with PsA, 8 with Ps, and 8 healthy controls. RESULTS: SHC levels were significantly higher in patients with RA (p = 0.01), as well as in those with PsA (p = 0.005) and Ps (p = 0.002) in comparison with the control group. However, a significant reduction in the TAC values in the serum of all three groups (RA, p = 0.03; PsA, p = 0.005; Ps, p = 0.001) were observed in comparison with the healthy controls. The thiolic concentration were found to have significantly diminished in patients with RA (p =0.0005) and Ps (p = 0.0005) in comparison with the control group. Our findings have brought out the fact that the therapeutic treatment of RA using biological drugs is more than satisfactory in accord with the considerable increase in the TAC values, although not significantly, compared to those patients treated with DMARDs. CONCLUSIONS: The determination of the parameters of oxidative stress utilising these methods may be useful as a quick test, and as routine in monitoring the state of oxidative stress in patients suffering from RA, PsA, and Ps, so that a more effective treatment for ROS can be undertaken accordingly. The administration of biological drugs seems to have a role in increasing the mechanism of the barrier which the body possesses against oxidative stress.

Definition of adult-onset rheumatoid arthritis from elderly-onset rheumatoid arthritis by studying T-lymphocyte subpopulations, their soluble receptors and soluble receptor of interleukin-2.

AIMS: The study of the distribution of T-lymphocyte sub-populations has revealed some immune characteristics of rheumatoid arthritis (RA) as well as polymyalgia rheumatica (PMR). There is much evidence that the subsets of T-lymphocyte subpopulations are well correlated with the age of the patient and the precise diagnosis of RA and PMR. The aims of the study were to evaluate the absolute number and percentage of T-lymphocyte subpopulation subsets in peripheral blood and their soluble receptors and serum soluble receptors of interleukin-2. MATERIAL AND METHODS: Thirty-six patients with RA were divided into 21 adult-onset RA (AoRA) and 15 elderly-onsets RA (EoRA) patients. They were compared with 48 PMR patients, 21 normal subjects under 45 years and 17 healthy elderly subjects over 65 years. T-lymphocyte subsets were studied by FACSCAN with double stained specific monoclonal antibodies. The EL ISA method was used to determine soluble receptors of CD4+ and CD8+ and IL-2. RESULTS: The AoRA patients had a significant alteration of T-lymphocyte sub-populations as well as their specific soluble receptors compared to EoRA patients. On the other hand, distribution of T-lymphocyte sub-populations in EoRA patients was quite similar to that in PMR patients. CONCLUSIONS: This method is probably not applicable for daily routine clinical practice but provides some interesting data for differential diagnosis between RA and PMR.

[Osteoporosis and anorexia nervosa: ultrasonometric and biochemical evaluation--preliminary data]. / Osteoporosi e anoressia nervosa: valutazione ultrasonometrica e biochimica--dati preliminari.

AIMS: To evaluate the relationship between Anorexia Nervosa (AN) and Osteoporosis (OP) by bone heel ultrasonometric measurement and biochemical bone metabolic data. MATERIALS AND METHODS: AN, 15 patients (2 males / 13 females; mean age 27.5 yr, range 16-44; mean BMI 15.78, range 13-19.3); normal subjects (NS), 10 (2 males / 8 females; mean age 27.5 yr, range 17-40; mean BMI 26.2, range 25-28). An Hologic Sahara ultrasound heel instrument has been utilized in order to obtain the following parameters: broadband ultrasound attenuation (BUA), speed of sound (SOS), Stiffness' index, bone mineral density (BMD) and T-score. As markers of bone formation osteocalcin (OC, ng/ml) and of resorption pyridinolines (Pyr, pmol/umol creatinine e deoxy-Pyr) have been studied by standardized analytical methods. RESULTS: (expressed as x+/-SD). PATIENTS: BUA, 65+/-11.22 (p<0.01); SOS, 1544.14+/-73.5 (ns); Stiffness, 89.8+/-19.4 (p<0.01): BMD, 0.55+/-0.53 (p<0.01); T-score, -1.4+/-1.12 (p<0.01); OC, 4.05+/-2.3 (p<0.01); Pyr, 53+/-21 (ns); d-Pyr, 7.17+/-4.5 (ns). NS: BUA, 88.57+/-8.63; SOS, 1567.72+/-11.88; Stiffness, 108.07+/-4.97; BMD, 0.611+/-0.027; T-score, 0.22+/-0.3; OC, 8.5+/-4.5; Pyr, 60+/-25; d-Pyr, 8.5+/-3.5. CONCLUSIONS: Our data seem to confirm that AN represents an important risk factor for OP. The ultrasonometric data in AN patients document some statistically significant differences from SN in term of BMD and T-score reduction. The metabolic data show that OC is reduced in AN patients, on the contrary, no differences appear in term of resorption bone markers.

[Study of lymphocyte subpopulations in chronic autoimmune inflammatory rheumatic diseases. II. Polymyalgia rheumatica: behaviour of lymphocyte subpopulations during treatment with two different corticosteroids]. / Studio delle sottopopolazioni linfocitarie nelle connettiviti. II. La polimialgia reumatica: comportamento delle sottopopolazioni linfocitarie durante terapia con due diversi corticosteroidi.

AIM: The aim of this paper was to study the distribution of CD8+ subsets, of soluble receptors of CD8 and IL-2 during a steroid treatment with prednisone and deflazacort. METHODS: Forty-eight patients (9 males and 39 females, mean age 69.4(6.5 years) with active polymyalgia rheumatica (ESR 74 (18 mm, 1(st) h) were studied. In order to determine the distribution of lymphocyte subpopulations a panel of monoclonal antibodies was utilised. Flow cytometry with a FACSCAN machine and ELISA method were utilized. RESULTS: At base-time in comparison with normal subjects: reduction (P<0.05) of CD4(-)/CD8(+) (356+/-112/mL vs 564(132/mL), due to reduction (P<0.001) of CD8(+)bright (224(86/mL vs 426+/-124/mL) and CD8(+)bright/CD57(-) (123+/-44/mL vs 256(58/mL); increasing (P<0.001) of sCD8 (514+/-123 U/mL vs 312+/-102 U/mL) and (P<0.005) sIL-2r (984(346 U/mL vs 244+/-58 U/mL). Group-PDN: significant (P<0.001) reduction of CD4(-)/CD8(+) (466+/-102), CD89(+))bright (302(74), CD8+bright/CD57- (186+/-51), sCD8 (418+/-96) and of sIL-2r (450+/-163) at 1(st) week, and toward the normal range at 1(st) month. Group-DFZ: normal values at 6th month: CD4(-)/CD8(+) (497+/-133), CD8(+)bright (401+/-98), CD8(+)bright/CD57(-) (240+/-64), sCD8 (317+/-82), while sIL-2r è (P<0.0005) higher vs group-PDN. Group-PDN: VES <50 at 1st week, normal value (14+/-7) at 3(rd) month; PCR, 2.2+/-1.2 at 3(rd) month and 1(0.8 at 6th month. Group-DFZ: VES >20 (24+/-5) at 6th month e PCR increased. CONCLUSIONS: PDN shows a faster action vs DFZ. DFZ does not seem to be able in reducing sIL-2r probably showing a persistent inflammation and immune activation status.

Double modulation of 5-fluorouracil by methotrexate and high-dose L-leucovorin in advanced colorectal cancer.

A phase II trial was performed to evaluate the efficacy and toxicity of a double modulation of 5-fluorouracil (5-FU) by methotrexate (MTX) and L-leucovorin (L-LV) in patients with advanced recurrent (inoperable) or metastatic colorectal carcinoma (ACC). Between July 1993 and October 1995, 41 patients with ACC received a regimen that consisted of MTX 150 mg/m2 i.v., infused over a 20-minute period at hour 0, followed 19 hours later by L-LV 250 mg/m2 in a 2-hour i.v. infusion. 5-FU, 900 mg/m2, was administered by i.v. push injection at hour 20. Beginning 24 hours after MTX administration, all patients received four doses of L-LV, 15 mg/m2 i.m., every 6 hours. Cycles were repeated every 15 days. Two patients were not assessable for response. Objective regression was observed in 11 of 39 (28%) patients, [95% confidence interval (CI), 14-42%]. One (2%) patient achieved complete response (CR) and 10 (26%) partial response (PR). No change was recorded in 15 (39%) patients and progressive disease was noted in 13 (33%) patients. The median time to treatment failure was 6 months and the median survival time was 10 months. Toxicity was within acceptable limits, but one therapy-related death due to severe leukopenia was observed. The dose-limiting toxicity was mucositis. Eight episodes of grade 3 or 4 stomatitis were observed, and were responsible for dosage modifications of MTX and 5-FU. In conclusion, further in experimental and clinical studies are clearly necessary in order to design the best modulatory strategy of 5-FU.

Ifosfamide and vinorelbine as first-line chemotherapy for advanced non-small cell lung carcinoma.

We evaluated the efficacy and toxicity of the novel combination of ifosfamide (IFX) and vinorelbine (VNB) as first-line chemotherapy in patients with stage IIIB and IV non-small cell lung cancer (NSCLC). Between March 1993 and November 1994, 44 patients (17 stage IIIB; 27 stage IV) received a regimen consisting of IFX, 2 g/m2 in a 1-h infusion, days 1-3; mesna, 400 mg/m2 in an i.v. bolus at hours 0 and 4 and 800 mg orally at hour 8, days 1-3; and VNB, 35 mg/ m2 in a 20-min infusion, days 1 and 15. During the first course only, a half dose of VNB (17.5 mg/m2) was administered on days 8 and 22. Courses were repeated every 28 days. Forty patients were fully evaluable for response, and 44 were assessable for toxicity. Objective regression was recorded in 13 of 40 patients (33%). No patient achieved a complete response. Thirteen patients presented a partial response (33%); 17 (42%) had no change; and progressive disease was observed in 10 (25%). The median duration of response was 10 months, and the median time to treatment failure for the whole group was 4 months. Median survival was 11 months. The dose-limiting toxic effect was myelosuppression. Leukopenia occurred in 25 patients (57%) and was grade 3 or 4 in 8 patients (18%). Twelve patients (27%) developed peripheral neurotoxicity, while five had mild IFX-induced CNS toxicity. Phlebitis was observed in 15 of 30 patients (50%) who did not have central implantable venous systems. The IFX-VNB combination exhibited an activity against NSCLC that was among the highest reported for non-cisplatin-containing regimens, with a toxicity profile that was easily managed.

Ifosfamide and vinorelbine as first-line chemotherapy for metastatic breast cancer.

PURPOSE: To evaluate the efficacy and toxicity of the combination of ifosfamide (IFX) and vinorelbine (VNB) as first-line chemotherapy in metastatic breast cancer (MBC). PATIENTS AND METHODS: Between August 1993 and August 1995, 45 patients with untreated MBC received a regimen that consisted of IFX 2 g/m2 by 1-hour intravenous (i.v.) infusion on days 1 to 3, mesna 400 mg/m2 by i.v. bolus at hours 0 and 4 and 800 mg/m2 orally at hour 8 on days 1 to 3, and VNB 35 mg/m2 by 20-minute i.v. infusion on days 1 and 15. Courses were repeated every 28 days. During the first course only, half-dose VNB (17.5 mg/m2) was administered on days 8 and 22. The median age was 53 years and 30 patients (67%) were postmenopausal. Dominant sites of disease were soft tissue in nine patients, bone in seven, and visceral in 29. RESULTS: Objective responses (ORs) were recorded in 25 of 43 assessable patients (58%; 95% confidence interval, 43% to 73%). Complete remissions (CRs) occurred in six patients (14%) and partial remissions (PRs) in 19 (44%). No change (NC) was recorded in 10 patients (23%) and progressive disease (PD) in eight patients (19%). The median time to treatment failure was 12 months and the median survival duration 19 months. Myelosuppression was the limiting toxicity, mainly leukopenia in 32 patients (74%). In contrast, anemia and thrombocytopenia were mild. Other significant toxicities included peripheral neuropathy in nine patients (21%), constipation in 15 (35%), and myalgias in 11 (26%). CONCLUSION: IFX/VNB is an active combination against MBC with moderate toxicity and deserves further evaluation.